Skip to main content
SpringerLink
Log in

Characterization and Isolation of Very Small Embryonic-like (VSEL) Stem Cells Obtained from Various Human Hematopoietic Cell Sources

  • Published:
Stem Cell Reviews and Reports Aims and scope Submit manuscript

Abstract

Stem cell transplantation is one of the available treatments for leukemia, lymphoma, hereditary blood diseases and bone marrow failure. Bone marrow (BM), peripheral blood progenitor cells (PBPC), and cord blood (CB) are the predominant sources of stem cells. Recently a new type of stem cell with a pluripotent potential has been identified. These cells were named “very small embryonic like stem cells (VSELs)”. It is claimed that VSEL stem cells can be found in adult BM, peripheral blood (PB), CB and other body tissues. This study is designed to characterize and isolate VSEL stem cells from different human hematopoietic sources; CB, PB and apheresis material (PBPC). VSEL stem cells were isolated from MNC and erythrocyte layers for all materials by using centrifugation and ficoll gradient method. We determined embryonic markers by flow cytometry, immunofluorescence and western blotting methods. Results from western blotting and immunofluorescence show high level of NANOG and OCT4 protein expression in PB, apheresis material and CB. Immunofluorescence images showed cytoplasmic and nuclear presence of these proteins. Flow cytometry results exhibited a higher expression of VSELs markers on debris area than CD45- population and higher expression on CB than PB. As a result, these findings have shown that it is necessary to investigate the function of pluripotent stem cell markers in differentiated adult cells. We further conclude that erythrocyte lysis method had the highest cell recovery amount among erythrocyte lysis and ficoll gradient methods. Consequently, this study gives us new information and viewpoints about expression of pluripotent stem cell (PSC) markers in adult tissues.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Perseghin, P. (2012). Stem cell therapy, regenerative medicine and hematopoietic stem cell transplantation: Recent achievements. Transfusion and Apheresis Science, 47(2), 191–192.

    Article  PubMed  Google Scholar 

  2. Lu, H., Xie, C., Zhao, Y. M., & Chen, F. M. (2013). Translational research and therapeutic applications of stem cell transplantation in periodontal regenerative medicine. Cell Transplantation, 22(2), 205–229.

    Article  PubMed  Google Scholar 

  3. Mamidi, M. K., Dutta, S., Bhonde, R., Das, A. K., & Pal, R. (2014). Allogeneic and autologous mode of stem cell transplantation in regenerative medicine: Which way to go? Medical Hypotheses, 83(6), 787–791.

    Article  CAS  PubMed  Google Scholar 

  4. Chivu-Economescu, M., & Rubach, M. (2017). Hematopoietic stem cells therapies. Current Stem Cell Research & Therapy, 12(2), 124–133.

    Article  CAS  Google Scholar 

  5. Barriga, F., Ramirez, P., Wietstruck, A., & Rojas, N. (2012). Hematopoietic stem cell transplantation: Clinical use and perspectives. Biological Research, 45, 307–316.

    Article  CAS  PubMed  Google Scholar 

  6. Kaufman, D. S. (2009). Toward clinical therapies using hematopoietic cells derived from human pluripotent stem cells. Blood, 114(17), 3513–3521.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Perutelli, P., Catellani, S., Scarso, L., Cornaglia-Ferraris, P., & Dini, G. (1999). Processing of human cord blood by three different procedures for red blood cell depletion and mononuclear cell recovery. Vox Sanguinis, 76, 237–240.

    Article  CAS  PubMed  Google Scholar 

  8. Rocha, V., Gluckman, E., & Eurocord and European Blood and Marrow Transplant Group. (2006). Clinical use of umbilical cord blood hematopoietic stem cells. Biology of Blood and Marrow Transplantation, 12, 34–41.

    Article  PubMed  Google Scholar 

  9. Kucia, M., Reca, R., Campbell, F. R., Zuba-Surma, E., Majka, M., Ratajcsak, J., et al. (2006). A population of very embryonic-like (VSEL) CXCR4+ SSEA-1+ OCT 4+ stem cells identified in adult bone marrow. Leukemia, 20, 857–869.

    Article  CAS  PubMed  Google Scholar 

  10. Kucia, M., Halasa, M., Wysoczynski, M., Baskiewicz-Masiuk, M., Moldenhawer, S., Zuba-Surma, E., Czajka, R., Wojakowski, W., Machalinski, B., & Ratajczak, M. Z. (2007). Morphological and molecular characterization of novel population of CXCR4+/SSEA-4+/Oct-4+ very small embryonic-like cells purified from human cord blood – Preliminary report. Leukemia, 21, 297–303.

    Article  CAS  PubMed  Google Scholar 

  11. Zuba-Surma, E. K., Kucia, M., Ratajczak, J., & Patajczak, M. Z. (2009). “Small stem cells” in adult tissues: Very small embryonic-like stem cells (VSELs) stand up. Cytometry, 75(1), 4–13.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Ratajczak, M. Z., Zuba-Surma, E., Wojaskowski, W., Suszynzka, M., Liu, R., Ratajczak, J., et al. (2014). Very small embryonic-like stem cells (VSELs) represent a real challenge in stem cell biology: Recent pros and cons in the midst of a lively debate. Leukemia, 28, 473–484.

    Article  CAS  PubMed  Google Scholar 

  13. Parker, G. C. (2014). Very small embryonic-like stem cells: A scientific debate? Stem Cells and Development, 23(7), 687–688.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Szade, K., Bukowska-Strakova, K., Nowak, W. N., Szade, A., Kachamakova-Trojanowska, N., Zukowska, M., Jozkowicz, A., & Dulak, J. (2013). Murine bone marrow Lin-Sca-1+CD45- very small embryonic-like (VSEL) cells are heterogeneous population lacking Oct-4A expression. PLoS One, 8(5), e63329.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Kucia, M., Wysoczynski, M., Wu, W., Zuba-Surma, E. K., Ratajczak, J., & Ratajczak, M. Z. (2008). Evidence that very small embryonic-like stem cells are mobilized into peripheral blood. Stem Cells, 26, 2083–2092.

    Article  CAS  PubMed  Google Scholar 

  16. Sovalat, H., Scrofani, M., Eidenschenk, A., Pasquet, S., Rimelen, V., & Henon, P. (2011). Identification and isolation from either adult human bone marrow or G-CSF-mobilized peripheral blood of CD34+/CD133+/CXCR4+/ Lin-CD45- cells, featuring morphological, molecular, and phenotypic characteristics of very small embryonic-like (VSEL) stem cells. Experimental Hematology, 39, 495–505.

    CAS  PubMed  Google Scholar 

  17. Halasa, M., Baskiewicz-Masiuk, M., Dabrkomska, E., & Machalinski, B. (2008). An efficient two-step method to purify very small embryonic-like (VSEL) stem cells from umbilical cord blood. Folia Histochemical Cytobiology, 46(2), 239–243.

    Google Scholar 

  18. Loos, H., Blok-Schut, B., Van Doorn, R., Hoksbergen, R., Riviere, A. B., & Meerhof, L. (1976). A method for the recognition and separation of human blood monocytes on density gradient. Blood, 48, 731–742.

    Article  CAS  Google Scholar 

  19. Bhartiya, D., Shaikh, A., Nagvenkar, P., Kasiviswananthan, S., Pethe, P., Pawani, H., et al. (2012). Very small embryonic-like stem cells with maximum regenerative potential get discarded during cord blood banking and bone marrow processing for autologous stem cell therapy. Stem Cell and Development, 21(1).

  20. Chang, Y., Tien, K., Wen, C., Hsieh, T., & Hwang, S. (2014). Recovery of CD45(−)/Lin(−)/SSEA-4(+) very small embryonic-like stem cells by cord blood bank standard operating procedures. Cytotherapy, 0, 1–6.

    Google Scholar 

  21. Sovalat, H., Scrofani, M., Eidenschenk, A., Pasquet, S., Rimelen, V., & Henon, P. (2011). Identification and isolation from either adult human bone marrow or G-CSF-mobilized peripheral blood of CD34+/CD133+/CXCR4+/ Lin-CD45- cells, featuring morphological, molecular, and phenotypic characteristics of very small embryonic-like (VSEL) stem cells. Experimental Hematology, 39, 495–505.

    CAS  PubMed  Google Scholar 

  22. Ratajczak, M. Z., Zuba-Surma, E., Wojakowski, W., Kucia, M., & Ratajczak, J. (2008). Bone marrow – Home of versatile stem cells. Transfusion Medicine and Hemotherapy, 35, 248–259.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Kucia, M., Reca, R., Jala, V. R., Dawn, B., Ratajczak, J., & Ratajczak, M. Z. (2005). Bone marrow as a home of heterogenous population of nonhematopoietic stem cells. Leukemia, 19, 1118–1127.

    Article  CAS  PubMed  Google Scholar 

  24. Kucia, M., Ratajczak, J., & Ratajczak, M. Z. (2005). Are bone marrow stem cells plastic or heterogenous – That is the question. Experimental Hematology, 33, 613–623.

    Article  PubMed  Google Scholar 

  25. Lee, J., Kim, H. K., Rho, J., Han, Y., & Kim, J. (2006). Human Oct4 isoforms differ in their ability to confer self-renewal. The Journal of Biological Chemistry, 281, 33554–33565.

    Article  CAS  PubMed  Google Scholar 

  26. Zangrossi, S., Marabese, M., Broggini, M., Giordano, R., D’Erasmo, M., Montelatici, E., et al. (2007). Oct-4 expression in adult human differentiated cells challenges its role as a pure stem cell marker. Stem Cells, 25, 1675–1680.

    Article  CAS  PubMed  Google Scholar 

  27. Warthemann, R., Eildermann, K., Debowski, K., & Behr, R. (2012). False-positive antibody signals for the pluripotency factor OCT4A (POU5F1) in testis-derived cells may lead to erroneous data and misinterpretations. Molecular Human Reproduction, 18(12), 605–612.

  28. Wang, Y., & Teng, J. S. (2016). Increased multi-drug resistance and reduced apoptosis in osteosarcoma side population cells are crucial factors for tumor recurrence. Experimental and Therapeutic Medicine, 12(1), 81–86.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Fang, F., Angulo, B., Xia, N., Sukhwani, M., Wang, Z., Carey, C. C., Mazurie, A., Cui, J., Wilkinson, R., Wiedenheft, B., Irie, N., Surani, M. A., Orwig, K. E., & Reijo Pera, R. A. (2018). PAX5-OCT4-PRDM1 developmental switch specifies human primordial germ cells. Nature Cell Biology, 20(6), 655–665.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Ambady, S., Malcuit, C., Kashpur, O., Kole, D., Holmes, W. F., Hedblom, E., Page, R. L., & Dominko, T. (2010). Expression of NANOG and NANOGP8 in a variety of undifferentiated and differentiated human cells. The Int of Develop Biol., 54, 1743–1754.

    Article  CAS  Google Scholar 

  31. Booth, H. A. F., & Holland, P. W. H. (2004). Eleven daughters of NANOG. Genomics, 84, 229–238.

    Article  CAS  PubMed  Google Scholar 

  32. Levasseur, D. N., & Das, S. v. J. (2011). Alternative splicing produces nanog protein variants with different capacities for self-renewal and pluripotency in embriyonic stem cells. The Journal of Biological Chemistry, 286, 42690–42703.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Gu, T., Liu, S., & Zheng, P. (2012). Cytoplasmic NANOG- positive stromal cells promote human cervical cancer progression. The American Journal of Pathology, 181(2), 652–660.

    Article  CAS  PubMed  Google Scholar 

  34. Ezeh, U. I., Turek, P. J., Reijo, R. A., & Clark, A. T. (2005). Human embryonic stem cell genes OCT4, NANOG, STELLAR, and GDF3 are expressed in both seminoma and breast carcinoma. Cancer, 104, 2255–2265.

    Article  CAS  PubMed  Google Scholar 

  35. Page, R., Ambady, S., Holmes, W. F., Vilner, L., Kole, D., Kashpur, O., et al. (2009). Induction of stem cell gene expression in adult human fibroblasts without transgenes. Cloning and Stem Cells, 11(3), 417–425.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Alvarez, A., Hossain, M., Dantuma, E., Merchant, S., & Sugaya, K. (2010). Nanog overexpression allows human mesenchymal stem cells to differentiate into neural cells. Neuroscience and Medicine, 1, 1–13.

    Article  CAS  Google Scholar 

  37. Eberle, I., Pless, B., Braun, M., Dingermann, T., & Marschalek, R. (2010). Transcriptional properties of human NANOG1 and NANOG2 in acute leukemic cells. Nucleic Acids Research, 38(16), 5384–5395.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work supported by Scientific Research Projects Coordination Unit of Istanbul University. Project number 35553.

Author information

Authors and Affiliations

  1. Deparment of Physiology, Istanbul Medical Faculty, Istanbul University, Istanbul, Turkey

    Serap Erdem Kuruca & Dolay Damla Çelik

  2. Department of Genetic and Bioengineering, Faculty of Engineering and Architecture, Kastamonu University, Kastamonu, Turkey

    Dilşad Özerkan

  3. Deparment of Molecular Medicine, The Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey

    Gökçe Erdemir

Authors
  1. Serap Erdem Kuruca
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dolay Damla Çelik
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dilşad Özerkan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gökçe Erdemir
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Dilşad Özerkan.

Ethics declarations

Conflict of Interest

All authors declare no conflicts of interest.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kuruca, S.E., Çelik, D.D., Özerkan, D. et al. Characterization and Isolation of Very Small Embryonic-like (VSEL) Stem Cells Obtained from Various Human Hematopoietic Cell Sources. Stem Cell Rev and Rep 15, 730–742 (2019). https://doi.org/10.1007/s12015-019-09896-1

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12015-019-09896-1

Keywords

Search

Navigation